1. Pakistan Veterinary Journal
ISSN: 0253-8318 (PRINT), 2074-7764 (ONLINE)
Accessible at: www.pvj.com.pk
Haematological and Serum Protein Values in Tuberculin Reactor and Non-Reactor Water
Buffaloes, Cattle, Sheep and Goats
M. Tariq Javed*, L. Ahmad, M. Irfan, I. Ali, A. Khan, M. Wasiq, F. A. Farooqi, A. Latif Shahid and M. Cagiola2
Department of Pathology, Faculty of Veterinary Science, University of Agriculture Faisalabad, Pakistan; 2Instituto
Zooprofilattico Sperimentale dell'Umbria e delle Marche, Perugia, Italy
*Corresponding author: mtjaved_uaf@hotmail.com
ARTICLE HISTORY ABSTRACT
Received: October 25, 2009 The diagnostic accuracy of tuberculosis can be enhanced by new tests like gamma
Revised: January 11, 2010 interferon used in conjunction with tuberculin test but these tests are quite
Accepted: February 26, 2010 expensive to be conducted on all animals in resource poor countries. The present
Key words: study was conducted to clarify the usefulness of haematological parameters and
Buffalo changes in serum proteins along with tuberculin test in different animal species. In
Cattle buffaloes, significantly lower RBC count, WBC count, Hb concentration,
Goat neutrophils and eosinophils while significantly higher monocytes percentage along
Haematology with serum total proteins, albumin and globulins were observed in tuberculin
Serum proteins positive reactors. In cattle, significantly lower RBC count, PCV, neutrophils and
Sheep serum albumin, while lymphocyte percentage was significantly higher in positive
Tuberculosis reactor animals. In sheep, PCV was lower in positive reactors. In goats, basophils
percentage was significantly higher (P = 0.002) in positive reactors. A significant
positive correlation of Hb with RBC and PCV in positive reactor buffaloes, goats
and sheep was observed. The correlation statistics for differential leukocyte
percentage did not differ between positive and negative reactor animals of these
species. In cattle and goats, albumin showed significant positive correlation with
globulins in negatively reacting animals, while this correlation was non-significant
in positive reactor animals. In buffaloes, albumin showed a significant positive
correlation with globulins and this correlation was non-significant in negative
reactor buffaloes. It was concluded that tuberculosis caused decrease in erythrocyte
parameters including RBC count, PCV and Hb concentration, while it caused an
increase in monocytes, eosinophils and lymphocytes.
©2010 PVJ. All rights reserved
To cite this article: Javed MT, L Ahmad, M Irfan, I Ali, A Khan, M Wasiq, FA Farooqi, MS Latif and M Cagiola,
2010. Haematological and serum protein values in tuberculin reactor and non-reactor water buffaloes, cattle, sheep
and goats. Pak Vet J, 30(2): 100-104.
INTRODUCTION other organs (Menzies and Neill, 2000). The causative
organism not only produces tubercles but also affects
Bovine tuberculosis is one of the most important hemato-biochemical parameters as various diseases have
zoonotic diseases of cattle and other farm animals. It is an an adverse effect on the haematological parameters (Sattar
infectious disease caused by Mycobacterium bovis that and Mirza, 2009). According to Samad and Rahman
has a broad host range (Wedlock et al., 2002). This (1986), hemoglobin concentration, total erythrocytes and
disease has socio-economic or public health importance neutrophils significantly decrease whereas values of
and is of great significance to international trade of erythrocytes sedimentation rate, total leukocyte counts
animals and animal products (OIE, 2004). Not only, and lymphocytes significantly increase in positive reactors
disease poses a greater threat to the economies of than healthy cattle. Moreover, depending on the stage and
developing countries but also remains a problem in progress of the disease, leukocytosis, leukopenia,
developed world (Khan et al., 2008). It is a contagious lymphocytosis, lymphopenia, neutrophilia and monocyto-
chronic debilitating disease of animals associated with sis can occur (Kumar et al., 1994).
progressive emaciation and tubercle (granuloma) The diagnosis of this disease in routine is made by
formation that are mostly confined to respiratory system, intradermal tuberculin test, while the gold standard is
primarily in the lungs, lymph nodes and occasionally in histopathological examination of morbid tissues and
100

2. 101 Pak Vet J, 2010, 30(2): 100-104.
culture isolation of organism. Recently, new tests have separated in another glass tube without EDTA to collect
been developed like gamma interferon ELISA but these serum. The haematological studies were carried out as
tests are quite expensive to be conducted routinely in described by Benjamin (1978). Serum total proteins were
resource poor countries. These tests are reported to be determined by the Biuret method, as described by Oser
used in conjunction with tuberculin test for more accurate (1976). The serum albumin was determined following the
diagnosis. In the absence of these tests for poor countries, method of Varley et al. (1980). The globulin was estimated
the use of haemato-biochemical parameters could be of by subtracting serum albumin from serum total proteins.
value in enhancing the diagnostic accuracy. To have the The results obtained were subjected to statistical
idea of its usefulness in different animal species, the analysis by using General Linear Model procedure and
present work was conducted involving a larger sample means were compared by Dunett’s test on personal
size in buffaloes, cattle, sheep and goats along with computer using SAS statistical software (SAS, 2004).
comparative tuberculin test. Correlation statistics was also computed.
MATERIALS AND METHODS RESULTS
A total of 2057 buffaloes, 2273 cattle, 4983 sheep In buffaloes, the results on haematological parameters
and 1987 goats were tested using the single comparative showed significantly lower red blood cell count
intradermal tuberculin test at the neck region, using (P=0.0026), white blood cell count (P<0.0001),
bovine (50,000 IU/ml) and avian (25,000 IU/ml) PPDs haemoglobin (P<0.0001) concentration, neutrophils and
produced at the authorized laboratory at the Istituto eosinophils (P=0.01), while monocyte percentage was
Zooprofilattico Umbria e Marche, Italy. These animals higher in positive reactor than negative reactor animals
were from Government Livestock Experiment Stations of (Table 1). Serum total proteins, albumin and globulins
Punjab and villages of two districts of Punjab. Buffalo and were significantly (P<0.0001) higher in positive reactor
cattle studied in two cities were from the same locality as animals (Table 1).
for sheep and goats, while a total of 15 Government In cattle, haematological parameters revealed
livestock farms were included in the study. The negative significantly lower values of red blood cell count
animals included were from small holdings at villages and (P=0.005), packed cell volume (P<0.0001) and neutrophil
small number was taken from a farm where animals were percentage (P=0.0004), while lymphocyte percentage
newly purchased and were tested negative. The protocols (P=0.0003) was significantly higher in positive reactor
for the production of PPDs, the execution of the test and cattle (Table 2). Among serum proteins, only serum
its interpretation were carried out according to the criteria albumin was significantly lower (P=0.02) in positive
described in the OIE Manual of Standards for Diagnostic reactor cattle (Table 2).
Tests and Vaccines (OIE, 2004). In sheep, only packed cell volume (PCV) differed
Blood was collected from all the positive reactors significantly between positive and negative reactor
including, 137 buffaloes, 134 cattle, 36 sheep and 33 animals, while all other parameters of haematology and
goats. Almost same numbers of negative animals of about serum proteins did not differ statistically (Table 3). PCV
same status (age, parity etc.) were also included and blood percentage was lower (P=0.02) in positive reactor sheep
samples were collected from these animals (140 buffaloes, (Table 3). Similarly, haematological parameters and
130 cattle, 36 sheep and 33 goats). About 15 ml of whole serum proteins in goats did not differ statistically between
blood was collected from jugular vein of each animal. Out positive and negative reactor animals, except basophil
of this, 5 ml was separated in a clean glass tube having percentage which was higher (P=0.002) in positive reactor
sufficient quantity of EDTA and 10 ml of blood was animals (Table 4).
Table 1: Comparison of mean haematological parameters and serum proteins in tuberculin positive and negative
reactor buffaloes
Parameters studied Negative animals (n = 140) Positive reactors (n = 137) P value
RBC (106/µl) 5.77 ± 1.34 5.09 ± 1.81 0.002
WBC (103/µl) 5.88 ± 2.89 2.59 ± 3.44 <0.0001
Hb (g/dl) 13.86 ± 6.34 11.05 ± 2.07 <0.0001
PCV (%) 31.76 ± 4.59 32.91 ± 7.14
ESR (mm/hr) 79.54 ± 43.52 77.24 ± 44.17
Differential leukocyte count
Neutrophils (%) 40.82 ± 7.30 38.05 ± 8.62 0.01
Lymphocytes (%) 48.80 ± 7.30 50.62 ± 9.26
Monocytes (%) 3.92 ± 2.69 5.99 ± 3.35 <0.0001
Eosinophils (%) 6.40 ± 2.61 5.26 ± 3.99 0.01
Basophils (%) 0.27 ± 0.58 0.41 ± 1.11
Serum proteins
Serum total proteins (g/dl) 7.55 ± 1.61 10.18 ± 3.22 <0.0001
Serum albumin (g/dl) 3.77 ± 0.82 4.73 ± 1.60 <0.0001
Serum globulins (g/dl) 3.55 ± 0.88 5.07 ± 2.61 <0.0001

3. 102 Pak Vet J, 2010, 30(2): 100-104.
Table 2: Comparison of mean haematological parameters and serum proteins in tuberculin positive and negative
reactor cattle
Parameters studied Negative animals (n = 130) Positive reactors (n = 134) P value
RBC (106/µl) 6.04 ± 1.54 5.32 ± 1.51 0.005
WBC (103/µl) 6.26 ± 1.03 6.06 ± 1.71
Hb (g/dl) 9.71 ± 2.04 9.78 ± 1.71
PCV (%) 33.47 ± 2.91 30.56 ± 4.05 <.0001
ESR (mm/hr) 2.74 ± 8.30 2.80 ± 1.77
Differential leukocyte count
Neutrophils (%) 33.64 ± 7.97 27.57 ± 10.84 0.0004
Lymphocytes (%) 58.27 ± 6.97 64.00 ± 10.19 0.0003
Monocytes (%) 2.88 ± 1.19 2.89 ± 0.95
Eosinophils (%) 5.33 ± 1.70 5.22 ± 1.81
Basophils (%) 0.16 ± 0.42 0.30 ± 0.48
Serum proteins
Serum total proteins (g/dl) 11.93 ± 3.43 11.23 ± 2.94
Serum albumin (g/dl) 5.76 ± 1.13 5.17 ± 1.32 0.02
Serum globulins (g/dl) 5.97 ± 2.93 6.18 ± 2.58
Table 3: Comparison of mean haematological parameters and serum proteins in tuberculin positive and negative
reactor sheep
Parameters studied Negative animals (n = 36) Positive reactors (n = 36) P value
RBC (106/µl) 8.17 ± 1.28 7.83 ± 1.59
WBC (103/µl) 4.59 ± 2.11 8.47 ± 2.59
Hb (g/dl) 7.44 ± 3.25 8.05 ± 1.71
PCV (%) 26.80 ± 3.68 24.91 ± 2.95 0.02
ESR (mm/hr) 4.58 ± 8.34 2.30 ± 3.07
Differential leukocyte count
Neutrophils (%) 38.72 ± 7.23 37.94 ± 9.63
Lymphocytes (%) 38.32 ± 7.37 40.48 ± 8.68
Monocytes (%) 3.00 ± 1.97 3.57 ± 2.47
Eosinophils (%) 16.60 ± 11.07 15.57 ± 10.24
Basophils (%) 1.16 ± 1.72 1.45 ± 1.96
Serum proteins
Serum total proteins (g/dl) 10.31 ± 2.45 11.16 ± 3.51
Serum albumin (g/dl) 6.28 ± 2.36 6.23 ± 2.41
Serum globulins (g/dl) 3.99 ± 1.68 4.97 ± 2.17
Table 4: Comparison of mean haematological parameters and serum proteins in tuberculin positive and negative
reactor goats
Parameters studied Negative animals (n = 33) Positive reactors (n = 33) P value
RBC (106/µl) 8.52 ± 1.41 7.91 ± 1.34
WBC (103/µl) 6.47 ± 2.48 6.52 ± 2.39
Hb (g/dl) 7.83 ± 1.42 8.53 ± 1.32
PCV (%) 24.56 ± 3.74 23.96 ± 2.88
ESR (mm/hr) 1.40 ± 0.71 1.53 ± 0.96
Differential leukocyte count
Neutrophils (%) 38.78 ± 7.15 37.91 ± 7.61
Lymphocytes (%) 45.86 ± 10.17 43.89 ± 11.03
Monocytes (%) 3.17 ± 1.52 3.07 ± 2.30
Eosinophils (%) 12.04 ± 11.01 13.96 ± 10.50
Basophils (%) 0.41 ± 0.71 1.76 ± 1.56 0.002
Serum proteins
Serum total proteins (g/dl) 11.28 ± 6.65 9.68 ± 3.27
Serum albumin (g/dl) 6.19 ± 3.87 4.82 ± 1.94
Serum globulins (g/dl) 5.08 ± 3.24 4.86 ± 2.22

4. 103 Pak Vet J, 2010, 30(2): 100-104.
Correlation with severity, acuteness or chronicity of the disease.
The results of correlation statistics were different in Previously, a drop in red blood cell count and haemoglo-
positive and negative reacting animals of each species. A bin concentration (Samad and Rahman, 1986; Amin,
significant positive correlation of haemoglobin with red 1989; Rao et al., 1992; Kumar et al., 1994), PCV (Kumar
blood cell count and PCV in positive reacting buffaloes et al., 1994) and neutrophils (Samad and Rahman, 1986)
(r=0.348, P<0.0001; r=0.438, P<0.0001), goats (r=0.805, has been reported in positively reacting cattle. The
P<0.001; r=0.424, P =0.02) and sheep (r=0.731, increase in monocytes (in buffaloes) and lymphocytes (in
P<0.0001; r=0.353, P =0.03) was observed, while this cattle) correlated to some extent with earlier finding in
correlation was non-significant in negative animals of bison, where these were slightly higher than control
these species. The correlation statistics for differential animals (Miller et al., 1989). A significant decrease in
leukocyte percentages did not differ between positive and leukocytes in buffaloes, while non-significant decrease in
negative reactor animals of these species. cattle, was another supportive finding to tuberculin results
The correlation statistics of serum proteins revealed that has been previously reported and related with
similar findings in cattle and goats with albumin showing peracute to acute form of the disease (Lepper et al., 1977).
significant positive correlation with globulins in Similarly, increase in lymphocyte percentage and also
negatively reacting cattle (r=0.413, p=0.01) and goats total leukocyte count (contrary to our results) has been
(r=0.745, p<0.0001), while this correlation was non- reported (Samad and Rahman, 1986). Kumar et al. (1994)
significant in positive reactor animals of these species. also reported that depending on the stage and progress of
However, in buffaloes, albumin showed a significant the disease, leukocytosis, leucopoenia, lymphocytosis,
positive correlation with globulins (r=0.255, p=0.03), lymphopenia, neutrophilia and monocytosis can be
while this correlation was non-significant in negative observed. Our results also confirm the earlier findings
reactor buffaloes. (Kumar et al., 1994) about no utility of ESR in animals in
the diagnosis of tuberculosis or supportive diagnosis of
DISCUSSION tuberculin testing and thus our results disagree with the
increase in ESR in positive animals reported earlier
Haematological and serum protein variations in 18 (Samad and Rahman, 1986; Amin, 1989).
tuberculin positive buffaloes have previously been The results on these parameters in both sheep and
reported (Javed et al., 2006), while the data about goats were almost similar with non-significant difference
epidemiological factors in sheep and goats have been between positive and negative reactor animals, with
presented in another paper (Javed et al., 2010). This study exception of PCV which was significantly lower in sheep,
reports data of 137, 134, 36 and 33, positive reacting while non-significantly in goats. This suggests that these
buffaloes, cattle, sheep and goats, respectively to avian species behave differently than large ruminants. However,
and bovine PPD. Therefore, this data gives a better picture a significantly lower PCV in sheep was also observed in
about haematological and serum proteins variations positive cattle, thus supporting the results for this
among tuberculin positive and negative reacting animals parameter to be of some utility in sheep and cattle in
under tropical conditions. conjunction with results of tuberculin test.
The results of haematological and serum proteins The correlation statistics appears to have some more
parameters in buffaloes and cattle show species meaningful results in multi animal species. A significant
differences. However, some of the results were similar in positive correlation existed of Hb with RBC and PCV in
both species like the decrease in red blood cell count and three species of animals (buffaloes, goats and sheep),
neutrophil percentage. Positive reactor buffaloes showed while a significant negative correlation of albumin with
significant decrease in RBCs, WBCs, Hb concentration globulins was observed in two species (cattle and goats).
and eosinophils, while monocytes percentage along with The latter correlation in buffaloes was also significant but
serum total proteins, albumin and globulins showed positive. This suggests usefulness of the correlation
significant increase. However, in cattle, lymphocyte statistics along with haematology and serum proteins in
percentage was higher, while PCV and serum albumin strengthening the results of PPD testing in these species.
were lower in positive animals. This suggests a species This kind of data would be very helpful in the diagnosis of
variation. However, the results indicate that RBC count tuberculosis along with tuberculin testing in resource poor
and neutrophil percentage can be taken as subjective countries where advanced diagnostic facilities are costly
indicator along with results of PPD testing for and mostly unavailable and animal test and slaughter
confirmation of tuberculosis in these species. The policy along with strong abattoir vigilance is missing to
monocyte percentage in buffaloes and lymphocyte confirm the tuberculin results. Further, the results also
percentage in cattle can be considered supportive suggest that buffaloes behave differently than cattle and
indicators of disease in these species. There was however, these parameters (haematology and serum proteins) are
a clear variation in serum proteins pattern in both the probably of less significance in small ruminants.
species with increase in buffalo and decrease in cattle.
The results on studied parameters can however, vary with Conclusions
the stage of the disease in both the animal species (Lepper It can be concluded from the present study that
et al., 1977). We were unable to find any clinical sign tuberculosis causes decrease in erythrocyte parameters
indicative of tuberculosis in these animals nor we had any including RBC count, PCV and Hb concentration, while it
such history, therefore, experimental studies are needed to causes an increase in monocytes, eosinophils and
better correlate these results in both buffaloes and cattle lymphocytes. Further, the results of haemato- logy need to

5. 104 Pak Vet J, 2010, 30(2): 100-104.
be interpreted carefully in different animal species as Lepper AW, LA Comer and CW Pearson, 1977.
species variation does exist. Serological responses in experimental bovine
tuberculosis. Aust Vet J, 53: 301-305.
Acknowledgments Menzies FD and SD Neill, 2000. Cattle-to-cattle
Funding provided by the Pakistan Science transmission of bovine tuberculosis. Vet J, 160: 92-
Foundation, Islamabad under project grant PSF/Res/P- 106.
AU/AGR (283) and the bovine PPD donated by Istituto Miller LD, CO Thoen, KJ Throlson, EM Himes and LR
Zooprofi lattico Sperimentale dell'Umbria e delle Marche, Morgan, 1989. Serum biochemical and hematologic
Perugia, Italy is gratefully acknowledged. values of normal and Mycobacterium bovis infected
American Bison. J Vet Diagn Invest, 1: 219-222.
REFERENCES OIE, 2004. Manual of Standards for Diagnostic Tests and
Vaccines of World Organization for Animal Health
Amin S, 1989. Prevalence of buffalo tuberculosis by using (OIE), 4th Ed, http://www.oie.int/eng/normes/
short thermal test and identification of organism from mmanual/A_summary.htm.
lymph nodes. MSc Thesis, Department of Clinical Oser, BL, 1976. Hawk’s Physiological Chemistry.
Medicine and Surgery, College of Veterinary Mcgraw Hill Publ. Co New Delhi, India.
Science, Lahore, Pakistan. Rao, VNA, P Ramadas and M Dhinakran, 1992. A study
Benjamin MM, 1978. Outline of Veterinary Clinical on the effect of tuberculosis on body weight and
Pathology, 3rd Ed, The Iowa State Univ Press, Ames, haemogram values in cattle. Cheiron, 21: 19-22.
Iowa, USA. Samad MA and MS Rahman, 1986. Incidence of bovine
Khan IA, A Khan, A Mubarak and S Ali, 2008. Factors tuberculosis and its effect on certain blood indicies in
affecting prevalence of bovine tuberculosis in Nili dairy cattle of Bangladesh. Indian J Dairy Sci, 39:
Ravi buffaloes. Pak Vet J, 28(4): 155-158. 231-234.
Javed MT, M Usman, M Irfan and M Cagiola, 2006. A SAS, 2004. SAS Statistical Software Version 9.1. SAS
study on tuberculosis in buffaloes: some Institute Inc, Cary, NC, USA.
epidemiological aspects along with haematological Sattar A and RH Mirza, 2009. Haematological parameters
and serum protein changes. Vet Arhiv, 76: 193-206. in exotic cows during gestation and lactation under
subtropical conditions. Pak Vet J, 29(3): 129-132.
Javed MT, A Munir, M Shahid, G Severi, M Irfan, A
Varley H, AHG Owenlock and M Bell, 1980. Practical
Aranaz, and M Cagiola, 2010. Percentage of reactor
Clinical Biochemistry. Vol. 1. Williams and
animals to single comparative cervical intradermal
Heinmann Medical Books Ltd, London, UK, pp:
tuberculin (SCCIT) in small ruminants in Punjab
553–554.
Pakistan. Acta Tropica, 113: 88–91 Wedlock DN, MA Skinnee, GW De-Lisl and BM Buddle,
Kumar GS, PKR Lyer, MC Prasad and AK Sharma, 1994. 2002. Control of Mycpbacterium bovis infections and
Tuberculosis in cattle: Hematobiochemical studies. the risk to human population. Microb Infect, 4: 471-
Indian J Vet Path, 18: 38-42. 480.